Inactivation of thymidylate synthetase by a novel mechanism-based enzyme inhibitor: 1-(beta-D-2'-deoxyribofuranosyl) 8-azapurin-2-one 5'-monophosphate

The possible relationship of the soluble, “cytosolic” estradiol receptor with complex membranous and cytoskeletal structures of the cell matrix has been studied using a model erythrocyte system. Extraction of erythrocyte ghosts with a nonionic detergent (Triton X-100) under conditions that yield a cytoskeletal matrix reveals the presence of a limited number (less than 100) of specific sites having high affinity (k_d 10^?9 M) for the estradiol-receptor complex. The interation between the estradiol receptor and the cytoskeleton is critically dependent on temperature and it is improved by 25 mM KCl or NaCl and by 2.5 mM MgCl₂. The data suggest that the estradiol receptor, which has been generally considered to be freely “soluble” in the cytoplasm, may actually be physiologically associated in an integral manner with a complex cytoskeletal network in the cell cytoplasm.